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Tinospora cordifolia Induces Differentiation and Senescence Pathways in Neuroblastoma Cells

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Abstract

Children diagnosed with neuroblastomas often suffer from severe side as well as late effects of conventional treatments like chemotherapy and radiotherapy. Recent advances in understanding of molecular pathways involved in cellular differentiation and apoptosis have helped in the development of new therapeutic approach based on differentiation-based therapy of malignant tumours. Natural medicines with their holistic therapeutic approach are known to selectively eliminate cancer cells thus provide a better substitute for the conventional treatment modes. The current study was aimed to investigate the anti-cancer potential of aqueous ethanolic extract of Tinospora cordifolia (TCE) using IMR-32 human neuroblastoma cell line as a model system. TCE is highly recommended in Ayurveda for its general body and metal health-promoting properties. TCE treatment was seen to arrest the majority of cells in G0/G1 phase and modulated the expression of DNA clamp sliding protein (PCNA) and cyclin D1. Further, TCE-treated cells showed differentiation as revealed by their morphology and the expression of neuronal cell specific differentiation markers NF200, MAP-2 and NeuN in neuroblastoma cells. The differentiated phenotype was associated with induction of senescence and pro-apoptosis pathways by enhancing expression of senescence marker mortalin and Rel A subunit of nuclear factor kappa beta (NFkB) along with decreased expression of anti-apoptotic marker, Bcl-xl. TCE exhibited anti-metastatic activity and significantly reduced cell migration in the scratched area along with downregulation of neural cell adhesion molecule (NCAM) polysialylation and secretion of matrix metalloproteinases (MMPs). Our data suggest that crude extract or active phytochemicals from this plant may be a potential candidate for differentiation-based therapy of malignant neuroblastoma cells.

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Acknowledgments

The study was partially supported by University Grant Commission—University with Potential for Excellence (UGC-UPE, India) and Centre with Potential for Excellence in Particular Area (CPEPA UGC, India) grants to the University. Ms Rachana Mishra is thankful to the DBT, GOI for the research fellowship grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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The authors declare that no conflict of interests exists.

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  1. Department of Biotechnology, Guru Nanak Dev University, Amritsar, 143005, Punjab, India

    Rachana Mishra & Gurcharan Kaur

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  1. Rachana Mishra
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Correspondence to Gurcharan Kaur.

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Supplementary Fig. 1

(a) Flourescent images of MAP-2 immunostaining of primary hippocampal neuronal cells treated with TCE. (b) MTT assay for IMR-32 neuroblastoma cells treated with hexane and chloroform fractions of TCE. (c) LDH assay for IMR-32 cells treated with different concentration of hexane and chloroform fraction of TCE. (GIF 175 kb)

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Mishra, R., Kaur, G. Tinospora cordifolia Induces Differentiation and Senescence Pathways in Neuroblastoma Cells. Mol Neurobiol 52, 719–733 (2015). https://doi.org/10.1007/s12035-014-8892-5

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